Aim To explore the protective effect of lo-ganin ( an active component in Cornus officinalis ) on podocyte injury induced by advanced glycation end products ( AGEs) and its possible mechanism. Meth-ods Mouse podocytes were cultured in vitro and di-vided into Normal group, model group ( AGEs group) , loganin group and aminoguanidine group ( set as posi-tive control) . After being incubated with loganin( final concentrations are 0. 1, 1, 10 μmol · L-1 ) for 1 h, podocytes were stimulated by AGEs of 100 mg · L-1 for 24 h. Then, the cell viability was measured by u-sing MTT method. Podocytes apoptosis was evaluated by Hoechst33342/PI staining and flow cytometry. Re-ceptors of advanced glycation end products ( RAGE ) ,desmin and apoptosis-related protein like Bax, Bcl-2, cleaved caspase-3 in podocytes were detected by Western blot. Results Loganin ameliorated podocyte injury induced by AGEs, down-regulated the expression of desmin and RAGE. Loganin also reduced the apoptotic rate of podocytes and decreased the ratio of Bax/ Bcl-2 and the expression of pro-apoptotic protein cleaved caspase-3 in podocytes. Conclusion Loganin could ameliorate podocyte injury, and its mechanism may be related to the decrease of the expression of RAGE and inhibition of the apoptotic pathway.

BACKGROUND:The decel ularized porcine smal intestinal submucosa is a kind of bioactive extracel ular matrix, which is mainly composed of col agen, glycoprotein, proteoglycan and rich in col agen, glycosaminoglycan and various growth factors, and these components play an important role in promoting the differentiation and proliferation of tissue cel s. OBJECTIVE:To prepare the injectable smal intestinal submucosa and to investigate its co-culture with rat adipose-derived mesenchymal stem cel s in vitro. METHODS:The injectable smal intestinal submucosa and rat adipose-derived stem cel s were prepared. Cel counting kit-8 test for cel proliferation:Passage 3 adipose-derived stem cel s were seeded onto the injectable smal intestinal submucosa (experimental group) and cel s cultured under normal condition as control group. The cel proliferation was observed at 1, 3, 5 and 7 days of incubation. Live/dead staining test for the survival of cel s:Passage 3 adipose-derived stem cel s were respectively cultured in the injectable smal intestinal submucosa extracts (experimental group) and complete culture medium (control group). Cel survival was determined at 1, 3, 5 and 7 days of culture. RESULTS AND CONCLUSION:Scanning electron microscope oval and strip adipose-derived stem cel s adhered onto the material. The absorbance values in the experimental group were higher than those in the control group at 1 and 5 days of incubation (P<0.05). Cel survival:The number of cel s appeared to be in a rising trend with time in both two groups;after 1-day co-culture, al cel s in the two groups survived. Then dead cel s appeared in both two groups, showing no significant difference. These results show that the injectable smal intestinal submucosa exhibits a good cytocompatibility.

A colistin producing strain Paenibacillus polymyxa AS1.541 was treated by N-methyl-N-nitro-N-nitrosoguanidine(NTG) for increasing yields of the antibiotic colistin.High-yield strains were obtained by selection of deregulated mutant which grow on media containing colistin,a self second metabolite,and ethionine,an analogue of methionine.Some of these mutants have higher yield of colistin than that of the parent strain.

Objective To investigate the effect of down-regulated Blimp1 gene expression on differenetiation of bone marrow cells into dendritic cells (DCs).Methods Blimp1-shRNA was constructed and then loaded into lentivirus vector as lenti-blimp1-shRNA.Bone marrow cells from Balb/c mice were induced differentiation to DCs in an 8-day cell culture system with GM-CSF/IL-4 incubation and LPS stimulation at day 7.The cells were divided into groups as empty control (no treatment),lenti-control (transfected by lentivirus empty vector at day 1),and lenti-Blimp (transfected by lenti-blimp1-shRNA at day 1).The transfection efficiency was evaluated by GFP fluorescence for one week.The morphology and growth curve were analyzed.Real-time PT-PCR and Western blotting were used to evaluate mRNA and protein expression of Blimp1.At day 8,CD11 c and CD86/MHC-Ⅱ were quatitified using flow cytometry.Results GFP fluorescence emerged 3 days after transfection and was continuously expressed.Classic DC morphology was shown in no treatment cells,while damaged morphology presented in the cells with lentivirus transfection.The empty control cells proliferated from day 3,peaked as (2.45 ± 0.26) 106/well at day 4,and kept at (2.27 ± 0.19) 106/ well at day 8,The cells receiving lentivirus presented (1.69 ± 0.39) 106/well.The expression of Blimp1 mRNA and protein in the lenti-Blimp1 group was 76％/1％ and 1.0％/74.0％ of the empty control group.At day 8,CD11c,CD86 and MHC-Ⅱ expression in the empty control group was (69.2 ±5.0)％,(51.1± 4.9) ％ and (56.3 ± 7.3) ％,while (68.6±5.9)％,(49.5±4.3)％ and (69.4±4.5)％ in the lenti-control group,and (72.8 ± 5.5)％,(50.2 ± 6.0)％ and (46.5 ± 5.7)％ in the lenti Blimp1 group.Conclusion Lentivirus-mediated Blimp1-shRNA gene therapy modulates blimp1 expression of DC precursors.Down-regulation of Blimp1 fails to interrupt the differentiation of DCs but inhibits the maturation.

Objective To investigate the effect of adenosine A2A receptor on pituitary-adrenal axis response in acute phase of moderate craniocerebral trauma.Methods Eighteen adenosine A2A receptor knock-out mice in a C57BL/6 background and another eighteen their wild-type littermates were divided into normal control group and craniocerebral trauma for 4 hours group,and craniocerebral trauma for 24 hours group according to random number table,with siμ mice per group.Plasma levels of adrenocorticotropic-hormone (ACTH) and corticosterone at hours 4 and 24 postinjury were determined using ELISA method.Results At 4 and 24 hours,brain water content in wild-type mice [(80.950 ± 0.184) ％,(82.178 ± 0.255)％ respectively] was higher than that in gene knock-out mice [(80.006 ± 0.199)％,(81.091 ± 0.295)％ respectively,P ＜ 0.01].Besides,brain water content in both wild-type and gene knock-out mice increased after injury (P ＜ 0.01).Plasma levels of ACTH and corticosterone were higher in geneknock-out sham mice than in wild-type sham mice [(120.214 ± 2.472) ng/L vs (91.767 ±7.395) ng/L,(27.814 ±0.888) μg/L vs (11.430 ±0.644) μg/L respectively,P ＜0.0l].At 4 and 24 hours,plasma levels of ACTH [(174.776-± 5.040) ng/L,(189.613 ± 4.802) ng/L respectively] in geneknock-out mice showed a higher increase than those in wild-type mice [(119.594 ± 6.945) ng/L,(124.93-± 11.001 7) ng/L respectively,P ＜ 0.05].Moreover,plasma levels of corticosterone [(40.138 ±-0.805) μg/L] at 4 hours and [(37.440-0.485)μg/L] at 24 hours in gene knock-out mice showed a same result as compared with that in wild-type mice [(19.702 ± 0.804) μg/L,(17.602 ± 0.743) μg/L respectively,P ＜ 0.05].Conclusions Knock-out of adenosine A2A receptor increases the release of ACTH and corticosterone in acute stage of moderate craniocerebral trauma and promotes pituitary-adrenal stress response.This may provide a novel explanation for the neuroprotective effect of A2A receptor deficiency.

OBJECTIVE: To explore the relationship between best corrected visual acuity (BCVA) and refraction parameters in myopia. METHODS: Two thousand two hundred and seventy-four patients (4245 eyes) with different degrees of myopia were collected. Their BCVA, diopter of spherical (DS), diopter of cylinder (DC), astigmatism axis, axial length (AL) and corneal thickness were detected. The influence of those parameters on BCVA was studied and the mathematical model of the relationship between BCVA and other parameters including the age and gender of patients was established. RESULTS: The logistic regression analysis showed that there were correlations between the BCVA (y) and DS (x1), DC (x2), gender (x3), AL (x4), corneal thickness (x5), astigmatism axis (x6) and age (x7) (P<0.05): y=0.580 6-0.034 0 x1-0.046 8 x2+0.056 5 x3+0.016 5 x4+ 0.0007 x5+0.000 2 x6-0.005 8 x7. CONCLUSION For people with myopia, age, gender and corneal thickness have small effect on BCVA, while the DS, DC, AL and astigmatism axis have significant effect on BCVA. The BCVA would decline following the extension of DS, DC and AL. It is helpful to assess the vision of myopia by analyzing the refraction parameters comprehensively.
Adolescent ; Adult ; Child ; Cornea/pathology* ; Female ; Forensic Medicine/methods* ; Humans ; Male ; Middle Aged ; Models, Theoretical ; Myopia/physiopathology* ; Refraction, Ocular/physiology* ; Refractometry ; Visual Acuity ; Visual Fields/physiology* ; Young Adult

OBJECTIVETo investigate the changes of proto-oncogene c-fos, c-jun mRNA expression in angiotensin II (Ang II)-induced hypertrophy and effects of tanshinone II A (Tan) in the primary culture of neonatal rat cardiomyocytes.

METHODTwelve neonatal Wistar rats aged one day old of clean grade and both sexes were selected to isolate and culture cardiomyocytes. The cardiomyocytes were divided into: normal control group, Ang II (10(-6) mol x L(-1)) group, Ang II (10(-6) mol x L(-1)) +Tan (10(-8) g x L(-1)) group, Ang II (10(-6) mol x L(-1)) + valsartan (10(-6) mol x L(-1)) group, Tan (10(-8) g x L(-1)) group, valsartan (10(-6) mol x L(-1)) group. The cardiomyocyte size was determined by phase contrast microscope, the rate of protein synthesis in cardiomyocytes was measured by 3H-leucine incorporation. The c-fos, c-jun mRNA expression of cardiomyocytes were assessed using reverse transcription polymerase chain reaction (RT-PCR).

RESULTAng II was added to the culture medium and 30 min later, the c-fos, c-jun mRNA expression of cardiomyocytes increased significantly (P < 0. 01). After Ang II took effect for 24 h, the rate of protein synthesis in Ang II group increased more prominently than that in normal control group (P < 0.01). After Ang II took effect for 7 days, the size of cardiomyocyte in Ang II group increased obviously (P < 0. 05). If tanshinone II or valsartan was added to the culture medium before Ang II, both of them could inhibit the increase of c-fos, c-jun mRNA expression (P < 0.01), cardiomyocyte protein synthesis rate (P < 0.01), and cardiomyocyte size (P < 0.05) induced by Ang II.

CONCLUSIONTanshinone II could ameliorate Ang II-induced cardiomyocytes hypertrophy by inhabiting c-fos, c-jun mRNA expression.

Angiotensin II ; biosynthesis ; pharmacology ; Animals ; Cardiomegaly ; chemically induced ; metabolism ; pathology ; Diterpenes, Abietane ; Gene Expression Regulation ; drug effects ; Genes, fos ; genetics ; Genes, jun ; genetics ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Phenanthrenes ; pharmacology ; Proto-Oncogene Proteins c-fos ; genetics ; Proto-Oncogene Proteins c-jun ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Tetrazoles ; pharmacology ; Valine ; analogs & derivatives ; pharmacology ; Valsartan

OBJECTIVETo observe effects of tetrandrine (Tet) on angiotensin II (Ang II)-induced cardiomyocyte hypertrophy and the activity and expression of phosphorylated ERK1/2 (p-ERK1/2).

METHODIn the primary culture of neonatal rat cardiomyocytes, as indexes of cardiomyocyte hypertrophy, pulsation rate was measured under phase contrast microscope. Cell size was determined by cell morphology analytical system. The total protein was determined by coomassie brilliant blue and protein synthesis rate was measured by [3H]-Leucine incorporation. ERK activity was measured by immuno-precipitation. The expression of p-ERK1/2 was assessed using Western blot.

RESULTTet can decrease Ang II-induced elevations of the pulsation rate, cell size, total protein and protein synthesis rate; inhibit the activity and expression of p-ERK1/2.

CONCLUSIONThe anti-hypertrophic effect of Tet on Ang II-induced cardiomyocyte hypertrophy was associated with inhibition of ERK1/2 signaling pathway.

Angiotensin II ; toxicity ; Animals ; Animals, Newborn ; Benzylisoquinolines ; isolation & purification ; pharmacology ; Blotting, Western ; Cell Size ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hypertrophy ; Immunoprecipitation ; Microscopy, Phase-Contrast ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Phosphorylation ; drug effects ; Plants, Medicinal ; chemistry ; Protein Biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Stephania tetrandra ; chemistry

To improve the reliability and credibility of genotyping hepatitis E virus (HEV) and to explore the possibility of unifying standards of HEV genotyping by designing HEV universal primers for amplification of a long genomic fragment of different HEV genotypes. A set of universal primers (HEVuPrimer) was designed based on conserved regions determined by alignment analysis of 82 HEV strains with complete genome in GenBank. HEVuPrimer was compared with a set of previously used primers (MXJ primers) for their sequence-matching to different HEV strains and applied to amplify HEV genomic fragments from HEV reference strains with known different genotypes and clinical serum samples with anti-HEV-IgM by RT-nPCR. HEV genotyping based on the fragments amplified with HEVuPrimer was compared and validated with that based on HEV full genome and fragments obtained with MXJ primers. HEV genotyping by the phylogenetic analysis supplemented with the percent of nucleotide identity of the HEVuPrimer-determined fragments showed good correspondence with that based on HEV full-length genome. In addition, HEVuPrimer was much better than MXJ primers in matching sequences of HEV strains available from GenBank, and was able to amplify all the reference HEV strains with different genotypes. Among 124 samples with anti-HEV-IgM, 60 were positive for HEV RNA determined by a 644bp amplicon of RT-nPCR with the HEVuPrimenr. All the positive isolates belonged to HEV genotype 4 with nucleotide homology of 80.0%-99.9%, and could be further divided into 4 subgenotypes. Moreover, a novel subtype was identified with 6 HEV strains isolated very recently. The RT-nPCR using the HEVuPrimer and phylogenetic analysis of the amplified region provided strong evidences for its feasibility in HEV genetic classification. Our data have new implication for the consensus of genotype classification of HEV.
DNA Primers ; genetics ; Genome, Viral ; genetics ; Genotype ; Hepatitis E virus ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

ObjectiveTo investigate the development trend of drinking water type of endemic fluorosis in Qinghai province, and to provide the basis for the prevention and treatment of the disease. MethodsIn 2009, six monitoring counties were chosen by using simple random sampling methods, all diseased villages of the six monitoring counties were classified into light, moderate and severe disease types according to water fluorine content on the historical data, and 1 village was respectively chosen from each type. In monitoring villages with improved water, 3 tap water and one source water samples were collected, respectively. Five water samples were collected randomly in water unimproved monitoring villages according to water well locations of east, west, south, north and center. The fluorine content in water and urine was determined according to the Standard Testing Methods for Drinking Water (GB/T 5750-2006). Children aged 8 to 12 were examined for dental fluorosis by Dean method.Clinical osteofluorosis of all the resident over the age of 16 was examined, 2 village of these counties were randomly selected, and clinically diagnosed patients with skeletal fluorosis were examined again by X-ray using Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008). Urine sample of 30 children aged 8 to 12 and of 20 adults over the age of 16 were randomly collected and urinary fluoride was determined by F-ion selective electrode method (WS/T 89-2006). ResultsImproving water projects had been implemented in 14 monitoring villages of the 18 villages in 6 counties, the rate of improved-water was 77.78％(14/18). Among the 14 projects, 5 improved-water projects ran normally, and 9 projects ran with intermittently water supply. Seventy-five water samples were tested, themean of water fluoride was 0.48 mg/L. The prevalence of dental fluorosis was 31.95％ (285/892), that of clinical skeletal fluorosis was 36.55％(1570/4295) and the X-ray detection rate of skeletal fluorosis was 25.64％ (20/78).Five hundred and seventy-one urine samples of children were determined, and geometric mean of urinary fluorine was 1.04 mg/L; 370 adult urine samples were determined, and geometric mean of urinary fluorine was 1.52 mg/L Conclusion Epidemic of drinking water type of endemic fluorosis is still serious in Qinghai province, and drinking water defluoride measures should be further strengthened and improved.